+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-15780 | |||||||||
---|---|---|---|---|---|---|---|---|---|---|
Title | 2C9, C5b9-CD59 structure | |||||||||
Map data | 3C9, C5b9-CD59 local res filtered map (full map) | |||||||||
Sample |
| |||||||||
Function / homology | Function and homology information negative regulation of activation of membrane attack complex / cell killing / Terminal pathway of complement / membrane attack complex / complement binding / regulation of complement-dependent cytotoxicity / other organism cell membrane / regulation of complement activation / Activation of C3 and C5 / negative regulation of macrophage chemotaxis ...negative regulation of activation of membrane attack complex / cell killing / Terminal pathway of complement / membrane attack complex / complement binding / regulation of complement-dependent cytotoxicity / other organism cell membrane / regulation of complement activation / Activation of C3 and C5 / negative regulation of macrophage chemotaxis / Cargo concentration in the ER / complement activation, alternative pathway / complement activation / chemokine activity / COPII-mediated vesicle transport / retinol binding / endopeptidase inhibitor activity / tertiary granule membrane / positive regulation of vascular endothelial growth factor production / specific granule membrane / COPI-mediated anterograde transport / complement activation, classical pathway / positive regulation of chemokine production / transport vesicle / endoplasmic reticulum-Golgi intermediate compartment membrane / Peptide ligand-binding receptors / Regulation of Complement cascade / ER to Golgi transport vesicle membrane / protein homooligomerization / chemotaxis / positive regulation of immune response / extracellular vesicle / blood coagulation / G alpha (i) signalling events / blood microparticle / killing of cells of another organism / in utero embryonic development / vesicle / cell surface receptor signaling pathway / immune response / inflammatory response / G protein-coupled receptor signaling pathway / external side of plasma membrane / Golgi membrane / signaling receptor binding / focal adhesion / innate immune response / Neutrophil degranulation / protein-containing complex binding / endoplasmic reticulum membrane / cell surface / extracellular space / extracellular exosome / extracellular region / membrane / plasma membrane Similarity search - Function | |||||||||
Biological species | Homo sapiens (human) / human (human) | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 3.3 Å | |||||||||
Authors | Couves EC / Gardner S / Bubeck D | |||||||||
Funding support | European Union, United Kingdom, 2 items
| |||||||||
Citation | Journal: Nat Commun / Year: 2023 Title: Structural basis for membrane attack complex inhibition by CD59. Authors: Emma C Couves / Scott Gardner / Tomas B Voisin / Jasmine K Bickel / Phillip J Stansfeld / Edward W Tate / Doryen Bubeck / Abstract: CD59 is an abundant immuno-regulatory receptor that protects human cells from damage during complement activation. Here we show how the receptor binds complement proteins C8 and C9 at the membrane to ...CD59 is an abundant immuno-regulatory receptor that protects human cells from damage during complement activation. Here we show how the receptor binds complement proteins C8 and C9 at the membrane to prevent insertion and polymerization of membrane attack complex (MAC) pores. We present cryo-electron microscopy structures of two inhibited MAC precursors known as C5b8 and C5b9. We discover that in both complexes, CD59 binds the pore-forming β-hairpins of C8 to form an intermolecular β-sheet that prevents membrane perforation. While bound to C8, CD59 deflects the cascading C9 β-hairpins, rerouting their trajectory into the membrane. Preventing insertion of C9 restricts structural transitions of subsequent monomers and indirectly halts MAC polymerization. We combine our structural data with cellular assays and molecular dynamics simulations to explain how the membrane environment impacts the dual roles of CD59 in controlling pore formation of MAC, and as a target of bacterial virulence factors which hijack CD59 to lyse human cells. | |||||||||
History |
|
-Structure visualization
Supplemental images |
---|
-Downloads & links
-EMDB archive
Map data | emd_15780.map.gz | 332.9 MB | EMDB map data format | |
---|---|---|---|---|
Header (meta data) | emd-15780-v30.xml emd-15780.xml | 27.9 KB 27.9 KB | Display Display | EMDB header |
FSC (resolution estimation) | emd_15780_fsc.xml | 19.1 KB | Display | FSC data file |
Images | emd_15780.png | 54.2 KB | ||
Others | emd_15780_half_map_1.map.gz emd_15780_half_map_2.map.gz | 482.9 MB 482.8 MB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-15780 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-15780 | HTTPS FTP |
-Related structure data
Related structure data | 8b0gMC 8b0fC 8b0hC M: atomic model generated by this map C: citing same article (ref.) |
---|---|
Similar structure data | Similarity search - Function & homologyF&H Search |
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
---|---|
Related items in Molecule of the Month |
-Map
File | Download / File: emd_15780.map.gz / Format: CCP4 / Size: 600.7 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Annotation | 3C9, C5b9-CD59 local res filtered map (full map) | ||||||||||||||||||||
Voxel size | X=Y=Z: 0.831 Å | ||||||||||||||||||||
Density |
| ||||||||||||||||||||
Symmetry | Space group: 1 | ||||||||||||||||||||
Details | EMDB XML:
|
-Supplemental data
-Half map: 3C9, C5b9-CD59 unfiltered half map (full map)
File | emd_15780_half_map_1.map | ||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Annotation | 3C9, C5b9-CD59 unfiltered half map (full map) | ||||||||||||
Projections & Slices |
| ||||||||||||
Density Histograms |
-Half map: 3C9, C5b9-CD59 unfiltered half map (full map)
File | emd_15780_half_map_2.map | ||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Annotation | 3C9, C5b9-CD59 unfiltered half map (full map) | ||||||||||||
Projections & Slices |
| ||||||||||||
Density Histograms |
-Sample components
-Entire : 2C9, CD59 inhibited MAC Complex
Entire | Name: 2C9, CD59 inhibited MAC Complex |
---|---|
Components |
|
-Supramolecule #1: 2C9, CD59 inhibited MAC Complex
Supramolecule | Name: 2C9, CD59 inhibited MAC Complex / type: complex / ID: 1 / Chimera: Yes / Parent: 0 / Macromolecule list: #1-#7 Details: Solved in a DOPC, MSP2N2 nanodisc with a myristolated cytotopic CD59. |
---|---|
Source (natural) | Organism: Homo sapiens (human) / Location in cell: Serum |
-Macromolecule #1: Complement C5
Macromolecule | Name: Complement C5 / type: protein_or_peptide / ID: 1 / Number of copies: 1 / Enantiomer: LEVO |
---|---|
Source (natural) | Organism: human (human) |
Molecular weight | Theoretical: 188.512094 KDa |
Sequence | String: MGLLGILCFL IFLGKTWGQE QTYVISAPKI FRVGASENIV IQVYGYTEAF DATISIKSYP DKKFSYSSGH VHLSSENKFQ NSAILTIQP KQLPGGQNPV SYVYLEVVSK HFSKSKRMPI TYDNGFLFIH TDKPVYTPDQ SVKVRVYSLN DDLKPAKRET V LTFIDPEG ...String: MGLLGILCFL IFLGKTWGQE QTYVISAPKI FRVGASENIV IQVYGYTEAF DATISIKSYP DKKFSYSSGH VHLSSENKFQ NSAILTIQP KQLPGGQNPV SYVYLEVVSK HFSKSKRMPI TYDNGFLFIH TDKPVYTPDQ SVKVRVYSLN DDLKPAKRET V LTFIDPEG SEVDMVEEID HIGIISFPDF KIPSNPRYGM WTIKAKYKED FSTTGTAYFE VKEYVLPHFS VSIEPEYNFI GY KNFKNFE ITIKARYFYN KVVTEADVYI TFGIREDLKD DQKEMMQTAM QNTMLINGIA QVTFDSETAV KELSYYSLED LNN KYLYIA VTVIESTGGF SEEAEIPGIK YVLSPYKLNL VATPLFLKPG IPYPIKVQVK DSLDQLVGGV PVTLNAQTID VNQE TSDLD PSKSVTRVDD GVASFVLNLP SGVTVLEFNV KTDAPDLPEE NQAREGYRAI AYSSLSQSYL YIDWTDNHKA LLVGE HLNI IVTPKSPYID KITHYNYLIL SKGKIIHFGT REKFSDASYQ SINIPVTQNM VPSSRLLVYY IVTGEQTAEL VSDSVW LNI EEKCGNQLQV HLSPDADAYS PGQTVSLNMA TGMDSWVALA AVDSAVYGVQ RGAKKPLERV FQFLEKSDLG CGAGGGL NN ANVFHLAGLT FLTNANADDS QENDEPCKEI LRPRRTLQKK IEEIAAKYKH SVVKKCCYDG ACVNNDETCE QRAARISL G PRCIKAFTEC CVVASQLRAN ISHKDMQLGR LHMKTLLPVS KPEIRSYFPE SWLWEVHLVP RRKQLQFALP DSLTTWEIQ GVGISNTGIC VADTVKAKVF KDVFLEMNIP YSVVRGEQIQ LKGTVYNYRT SGMQFCVKMS AVEGICTSES PVIDHQGTKS SKCVRQKVE GSSSHLVTFT VLPLEIGLHN INFSLETWFG KEILVKTLRV VPEGVKRESY SGVTLDPRGI YGTISRRKEF P YRIPLDLV PKTEIKRILS VKGLLVGEIL SAVLSQEGIN ILTHLPKGSA EAELMSVVPV FYVFHYLETG NHWNIFHSDP LI EKQKLKK KLKEGMLSIM SYRNADYSYS VWKGGSASTW LTAFALRVLG QVNKYVEQNQ NSICNSLLWL VENYQLDNGS FKE NSQYQP IKLQGTLPVE ARENSLYLTA FTVIGIRKAF DICPLVKIDT ALIKADNFLL ENTLPAQSTF TLAISAYALS LGDK THPQF RSIVSALKRE ALVKGNPPIY RFWKDNLQHK DSSVPNTGTA RMVETTAYAL LTSLNLKDIN YVNPVIKWLS EEQRY GGGF YSTQDTINAI EGLTEYSLLV KQLRLSMDID VSYKHKGALH NYKMTDKNFL GRPVEVLLND DLIVSTGFGS GLATVH VTT VVHKTSTSEE VCSFYLKIDT QDIEASHYRG YGNSDYKRIV ACASYKPSRE ESSSGSSHAV MDISLPTGIS ANEEDLK AL VEGVDQLFTD YQIKDGHVIL QLNSIPSSDF LCVRFRIFEL FEVGFLSPAT FTVYEYHRPD KQCTMFYSTS NIKIQKVC E GAACKCVEAD CGQMQEELDL TISAETRKQT ACKPEIAYAY KVSITSITVE NVFVKYKATL LDIYKTGEAV AEKDSEITF IKKVTCTNAE LVKGRQYLIM GKEALQIKYN FSFRYIYPLD SLTWIEYWPR DTTCSSCQAF LANLDEFAED IFLNGC |
-Macromolecule #2: Complement component C7
Macromolecule | Name: Complement component C7 / type: protein_or_peptide / ID: 2 / Number of copies: 1 / Enantiomer: LEVO |
---|---|
Source (natural) | Organism: human (human) |
Molecular weight | Theoretical: 93.625328 KDa |
Sequence | String: MKVISLFILV GFIGEFQSFS SASSPVNCQW DFYAPWSECN GCTKTQTRRR SVAVYGQYGG QPCVGNAFET QSCEPTRGCP TEEGCGERF RCFSGQCISK SLVCNGDSDC DEDSADEDRC EDSERRPSCD IDKPPPNIEL TGNGYNELTG QFRNRVINTK S FGGQCRKV ...String: MKVISLFILV GFIGEFQSFS SASSPVNCQW DFYAPWSECN GCTKTQTRRR SVAVYGQYGG QPCVGNAFET QSCEPTRGCP TEEGCGERF RCFSGQCISK SLVCNGDSDC DEDSADEDRC EDSERRPSCD IDKPPPNIEL TGNGYNELTG QFRNRVINTK S FGGQCRKV FSGDGKDFYR LSGNVLSYTF QVKINNDFNY EFYNSTWSYV KHTSTEHTSS SRKRSFFRSS SSSSRSYTSH TN EIHKGKS YQLLVVENTV EVAQFINNNP EFLQLAEPFW KELSHLPSLY DYSAYRRLID QYGTHYLQSG SLGGEYRVLF YVD SEKLKQ NDFNSVEEKK CKSSGWHFVV KFSSHGCKEL ENALKAASGT QNNVLRGEPF IRGGGAGFIS GLSYLELDNP AGNK RRYSA WAESVTNLPQ VIKQKLTPLY ELVKEVPCAS VKKLYLKWAL EEYLDEFDPC HCRPCQNGGL ATVEGTHCLC HCKPY TFGA ACEQGVLVGN QAGGVDGGWS CWSSWSPCVQ GKKTRSRECN NPPPSGGGRS CVGETTESTQ CEDEELEHLR LLEPHC FPL SLVPTEFCPS PPALKDGFVQ DEGTMFPVGK NVVYTCNEGY SLIGNPVARC GEDLRWLVGE MHCQKIACVL PVLMDGI QS HPQKPFYTVG EKVTVSCSGG MSLEGPSAFL CGSSLKWSPE MKNARCVQKE NPLTQAVPKC QRWEKLQNSR CVCKMPYE C GPSLDVCAQD ERSKRILPLT VCKMHVLHCQ GRNYTLTGRD SCTLPASAEK ACGACPLWGK CDAESSKCVC REASECEEE GFSICVEVNG KEQTMSECEA GALRCRGQSI SVTSIRPCAA ETQ |
-Macromolecule #3: Complement component C8 beta chain
Macromolecule | Name: Complement component C8 beta chain / type: protein_or_peptide / ID: 3 / Number of copies: 1 / Enantiomer: LEVO |
---|---|
Source (natural) | Organism: human (human) |
Molecular weight | Theoretical: 67.136891 KDa |
Sequence | String: MKNSRTWAWR APVELFLLCA ALGCLSLPGS RGERPHSFGS NAVNKSFAKS RQMRSVDVTL MPIDCELSSW SSWTTCDPCQ KKRYRYAYL LQPSQFHGEP CNFSDKEVED CVTNRPCRSQ VRCEGFVCAQ TGRCVNRRLL CNGDNDCGDQ SDEANCRRIY K KCQHEMDQ ...String: MKNSRTWAWR APVELFLLCA ALGCLSLPGS RGERPHSFGS NAVNKSFAKS RQMRSVDVTL MPIDCELSSW SSWTTCDPCQ KKRYRYAYL LQPSQFHGEP CNFSDKEVED CVTNRPCRSQ VRCEGFVCAQ TGRCVNRRLL CNGDNDCGDQ SDEANCRRIY K KCQHEMDQ YWGIGSLASG INLFTNSFEG PVLDHRYYAG GCSPHYILNT RFRKPYNVES YTPQTQGKYE FILKEYESYS DF ERNVTEK MASKSGFSFG FKIPGIFELG ISSQSDRGKH YIRRTKRFSH TKSVFLHARS DLEVAHYKLK PRSLMLHYEF LQR VKRLPL EYSYGEYRDL FRDFGTHYIT EAVLGGIYEY TLVMNKEAME RGDYTLNNVH ACAKNDFKIG GAIEEVYVSL GVSV GKCRG ILNEIKDRNK RDTMVEDLVV LVRGGASEHI TTLAYQELPT ADLMQEWGDA VQYNPAIIKV KVEPLYELVT ATDFA YSST VRQNMKQALE EFQKEVSSCH CAPCQGNGVP VLKGSRCDCI CPVGSQGLAC EVSYRKNTPI DGKWNCWSNW SSCSGR RKT RQRQCNNPPP QNGGSPCSGP ASETLDCS |
-Macromolecule #4: Complement component C8 alpha chain
Macromolecule | Name: Complement component C8 alpha chain / type: protein_or_peptide / ID: 4 / Number of copies: 1 / Enantiomer: LEVO |
---|---|
Source (natural) | Organism: human (human) |
Molecular weight | Theoretical: 65.239152 KDa |
Sequence | String: MFAVVFFILS LMTCQPGVTA QEKVNQRVRR AATPAAVTCQ LSNWSEWTDC FPCQDKKYRH RSLLQPNKFG GTICSGDIWD QASCSSSTT CVRQAQCGQD FQCKETGRCL KRHLVCNGDQ DCLDGSDEDD CEDVRAIDED CSQYEPIPGS QKAALGYNIL T QEDAQSVY ...String: MFAVVFFILS LMTCQPGVTA QEKVNQRVRR AATPAAVTCQ LSNWSEWTDC FPCQDKKYRH RSLLQPNKFG GTICSGDIWD QASCSSSTT CVRQAQCGQD FQCKETGRCL KRHLVCNGDQ DCLDGSDEDD CEDVRAIDED CSQYEPIPGS QKAALGYNIL T QEDAQSVY DASYYGGQCE TVYNGEWREL RYDSTCERLY YGDDEKYFRK PYNFLKYHFE ALADTGISSE FYDNANDLLS KV KKDKSDS FGVTIGIGPA GSPLLVGVGV SHSQDTSFLN ELNKYNEKKF IFTRIFTKVQ TAHFKMRKDD IMLDEGMLQS LME LPDQYN YGMYAKFIND YGTHYITSGS MGGIYEYILV IDKAKMESLG ITSRDITTCF GGSLGIQYED KINVGGGLSG DHCK KFGGG KTERARKAMA VEDIISRVRG GSSGWSGGLA QNRSTITYRS WGRSLKYNPV VIDFEMQPIH EVLRHTSLGP LEAKR QNLR RALDQYLMEF NACRCGPCFN NGVPILEGTS CRCQCRLGSL GAACEQTQTE GAKADGSWSC WSSWSVCRAG IQERRR ECD NPAPQNGGAS CPGRKVQTQA C |
-Macromolecule #5: Complement component C8 gamma chain
Macromolecule | Name: Complement component C8 gamma chain / type: protein_or_peptide / ID: 5 / Number of copies: 1 / Enantiomer: LEVO |
---|---|
Source (natural) | Organism: human (human) |
Molecular weight | Theoretical: 22.302424 KDa |
Sequence | String: MLPPGTATLL TLLLAAGSLG QKPQRPRRPA SPISTIQPKA NFDAQQFAGT WLLVAVGSAC RFLQEQGHRA EATTLHVAPQ GTAMAVSTF RKLDGICWQV RQLYGDTGVL GRFLLQARDA RGAVHVVVAE TDYQSFAVLY LERAGQLSVK LYARSLPVSD S VLSGFEQR ...String: MLPPGTATLL TLLLAAGSLG QKPQRPRRPA SPISTIQPKA NFDAQQFAGT WLLVAVGSAC RFLQEQGHRA EATTLHVAPQ GTAMAVSTF RKLDGICWQV RQLYGDTGVL GRFLLQARDA RGAVHVVVAE TDYQSFAVLY LERAGQLSVK LYARSLPVSD S VLSGFEQR VQEAHLTEDQ IFYFPKYGFC EAADQFHVLD EVRR |
-Macromolecule #6: Complement component C9
Macromolecule | Name: Complement component C9 / type: protein_or_peptide / ID: 6 / Number of copies: 3 / Enantiomer: LEVO |
---|---|
Source (natural) | Organism: human (human) |
Molecular weight | Theoretical: 63.252301 KDa |
Sequence | String: MSACRSFAVA ICILEISILT AQYTTSYDPE LTESSGSASH IDCRMSPWSE WSQCDPCLRQ MFRSRSIEVF GQFNGKRCTD AVGDRRQCV PTEPCEDAED DCGNDFQCST GRCIKMRLRC NGDNDCGDFS DEDDCESEPR PPCRDRVVEE SELARTAGYG I NILGMDPL ...String: MSACRSFAVA ICILEISILT AQYTTSYDPE LTESSGSASH IDCRMSPWSE WSQCDPCLRQ MFRSRSIEVF GQFNGKRCTD AVGDRRQCV PTEPCEDAED DCGNDFQCST GRCIKMRLRC NGDNDCGDFS DEDDCESEPR PPCRDRVVEE SELARTAGYG I NILGMDPL STPFDNEFYN GLCNRDRDGN TLTYYRRPWN VASLIYETKG EKNFRTEHYE EQIEAFKSII QEKTSNFNAA IS LKFTPTE TNKAEQCCEE TASSISLHGK GSFRFSYSKN ETYQLFLSYS SKKEKMFLHV KGEIHLGRFV MRNRDVVLTT TFV DDIKAL PTTYEKGEYF AFLETYGTHY SSSGSLGGLY ELIYVLDKAS MKRKGVELKD IKRCLGYHLD VSLAFSEISV GAEF NKDDC VKRGEGRAVN ITSENLIDDV VSLIRGGTRK YAFELKEKLL RGTVIDVTDF VNWASSINDA PVLISQKLSP IYNLV PVKM KNAHLKKQNL ERAIEDYINE FSVRKCHTCQ NGGTVILMDG KCLCACPFKF EGIACEISKQ KISEGLPALE FPNEK |
-Macromolecule #7: Complement component C6
Macromolecule | Name: Complement component C6 / type: protein_or_peptide / ID: 7 / Number of copies: 1 / Enantiomer: LEVO |
---|---|
Source (natural) | Organism: human (human) |
Molecular weight | Theoretical: 104.91818 KDa |
Sequence | String: MARRSVLYFI LLNALINKGQ ACFCDHYAWT QWTSCSKTCN SGTQSRHRQI VVDKYYQENF CEQICSKQET RECNWQRCPI NCLLGDFGP WSDCDPCIEK QSKVRSVLRP SQFGGQPCTA PLVAFQPCIP SKLCKIEEAD CKNKFRCDSG RCIARKLECN G ENDCGDNS ...String: MARRSVLYFI LLNALINKGQ ACFCDHYAWT QWTSCSKTCN SGTQSRHRQI VVDKYYQENF CEQICSKQET RECNWQRCPI NCLLGDFGP WSDCDPCIEK QSKVRSVLRP SQFGGQPCTA PLVAFQPCIP SKLCKIEEAD CKNKFRCDSG RCIARKLECN G ENDCGDNS DERDCGRTKA VCTRKYNPIP SVQLMGNGFH FLAGEPRGEV LDNSFTGGIC KTVKSSRTSN PYRVPANLEN VG FEVQTAE DDLKTDFYKD LTSLGHNENQ QGSFSSQGGS SFSVPIFYSS KRSENINHNS AFKQAIQASH KKDSSFIRIH KVM KVLNFT TKAKDLHLSD VFLKALNHLP LEYNSALYSR IFDDFGTHYF TSGSLGGVYD LLYQFSSEEL KNSGLTEEEA KHCV RIETK KRVLFAKKTK VEHRCTTNKL SEKHEGSFIQ GAEKSISLIR GGRSEYGAAL AWEKGSSGLE EKTFSEWLES VKENP AVID FELAPIVDLV RNIPCAVTKR NNLRKALQEY AAKFDPCQCA PCPNNGRPTL SGTECLCVCQ SGTYGENCEK QSPDYK SNA VDGQWGCWSS WSTCDATYKR SRTRECNNPA PQRGGKRCEG EKRQEEDCTF SIMENNGQPC INDDEEMKEV DLPEIEA DS GCPQPVPPEN GFIRNEKQLY LVGEDVEISC LTGFETVGYQ YFRCLPDGTW RQGDVECQRT ECIKPVVQEV LTITPFQR L YRIGESIELT CPKGFVVAGP SRYTCQGNSW TPPISNSLTC EKDTLTKLKG HCQLGQKQSG SECICMSPEE DCSHHSEDL CVFDTDSNDY FTSPACKFLA EKCLNNQQLH FLHIGSCQDG RQLEWGLERT RLSSNSTKKE SCGYDTCYDW EKCSASTSKC VCLLPPQCF KGGNQLYCVK MGSSTSEKTL NICEVGTIRC ANRKMEILHP GKCLA |
-Macromolecule #8: CD59 glycoprotein
Macromolecule | Name: CD59 glycoprotein / type: protein_or_peptide / ID: 8 / Number of copies: 1 / Enantiomer: LEVO |
---|---|
Source (natural) | Organism: Homo sapiens (human) |
Molecular weight | Theoretical: 9.101303 KDa |
Recombinant expression | Organism: Escherichia coli BL21 (bacteria) |
Sequence | String: MLQCYNCPNP TADCKTAVNC SSDFDACLIT KAGLQVYNKC WKFEHCNFND VTTRLRENEL TYYCCKKDLC NFNEQLEN |
-Experimental details
-Structure determination
Method | cryo EM |
---|---|
Processing | single particle reconstruction |
Aggregation state | particle |
-Sample preparation
Buffer | pH: 7.4 Component:
Details: 20 mM HEPES pH 7.4, 120 mM NaCl | |||||||||
---|---|---|---|---|---|---|---|---|---|---|
Grid | Model: Quantifoil R1.2/1.3 / Material: COPPER / Mesh: 300 / Support film - Material: GRAPHENE OXIDE / Support film - topology: CONTINUOUS / Pretreatment - Type: PLASMA CLEANING / Pretreatment - Time: 60 sec. / Pretreatment - Atmosphere: AIR | |||||||||
Vitrification | Cryogen name: ETHANE / Chamber humidity: 95 % / Chamber temperature: 296 K / Instrument: FEI VITROBOT MARK II | |||||||||
Details | C5b, C6, C7, C8 and C9 purified components were sources from CompTech. Myrisolated CD59 was a gift from R. Smith. Individual components were reacted together empty DOPC nanodiscs then purified by sucrose gradient centrifugation. |
-Electron microscopy
Microscope | FEI TITAN KRIOS |
---|---|
Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
Electron optics | C2 aperture diameter: 100.0 µm / Calibrated defocus max: 4.0 µm / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELDBright-field microscopy / Cs: 2.7 mm / Nominal defocus max: 0.9 µm / Nominal defocus min: 1.0 µm / Nominal magnification: 105000 |
Sample stage | Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER |
Image recording | Film or detector model: GATAN K3 (6k x 4k) / Number grids imaged: 4 / Number real images: 52838 / Average exposure time: 2.0 sec. / Average electron dose: 50.0 e/Å2 / Details: Collected over 4 separate data collections |
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |